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Materials and methods for producing geminivirus resistant plants

a technology of geminivirus and material, applied in the direction of viruses/bacteriophages, organic chemistry, peptide sources, etc., can solve the problems of $125 million in tomato production in florida, major limiting factors of transmission geminiviruses, and $125 million in losses, and achieve the effect of providing resistan

Inactive Publication Date: 2005-12-29
UNIV OF FLORIDA RES FOUNDATION INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008] The subject invention pertains to materials and methods for producing plants that are resistant to infection by geminiviruses and other related viruses. Methods of the invention comprise transforming a plant with a polynucleotide wherein when the polynucleotide is expressed in the plant, the transformed plant exhibits resistance to infection when challenged with a plant virus. In a preferred embodiment, a plant is transformed with a polynucleotide encoding a Rep protein or a mutated Rep protein derived from tomato mottle geminivirus or from tomato yellow leaf curl virus (TYLCV-Is). The methods of the invention can be used to provide resistance to viral infection in plants such as tomato and tobacco.

Problems solved by technology

Whitefly-transmitted geminiviruses have become a major limiting factor in tomato production in Florida, the Caribbean and much of Latin America.
Tomato production in Florida has suffered significant losses (estimated at $125 million in 1990-91) due to tomato mottle virus (ToMoV) infection, which first appeared in 1989.
Incidences of TYLCV-Is are increasing and economic losses were felt this past fall (1998).
Geminiviruses are very difficult to economically manage in fresh market tomatoes, and practically impossible to manage in processing tomatoes.
These costs are expected to increase significantly as growers' struggle to manage TYLCV-Is.
In Caribbean countries geminiviruses have caused many small and medium size tomato growers to go out of business due the increases in costs of production and crop losses.
The use of these methods are expensive and are often not an economically or horticulturally realistic alternative.
The least expensive and most practical control of whitefly-transmitted geminiviruses is the use of resistant cultivars.
At this time there are no commercially available resistant tomato cultivars for the geminiviruses native to the Western Hemisphere.
There are several cultivars available which have tolerance to TYLCV-Is, however the fruit size and the horticultural attributes of these cultivars are unsuitable for production in Florida.
There are no commercially available ToMoV-resistant tomato cultivars.
This linkage has significantly delayed development of resistant plants.
However, resistance was overcome with time.

Method used

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  • Materials and methods for producing geminivirus resistant plants
  • Materials and methods for producing geminivirus resistant plants
  • Materials and methods for producing geminivirus resistant plants

Examples

Experimental program
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Effect test

example 1

Field Evaluation of Transgenic Resistance to ToMoV

[0028] Advanced breeding lines, FL 7324 and FL 7613 were transformed with ToMoV Rep gene using standard Agrobacterium-mediated transformation techniques. A vector comprising a 35S CaMV (non-enhanced) promoter linked to the Rep gene was used in the transformation. Plant tissue was wounded using tungsten. Plants that contained a Rep transgene were identified using PCR methods. Those plants were then evaluated for resistance to viral infection. Untransformed parents and transformed lines from three different transformation events were evaluated for resistance to ToMoV and for yield, both in the presence and absence of ToMoV. Lines shown in the following tables are four generations past transformation. Lines 02 and 04 are from the same T0 plant in a FL 7613 background, lines 09 and 10 are from a second T0 plant and their background is FL 7324, and lines 11 and 12 are from a third T0 plant, and their background is FL 7613.

[0029] Souther...

example 2

Performance in the Presence of ToMoV and Whiteflies

[0030] For three seasons, Fall 1997, Spring 1998, and Fall 1998, tomato transplants to be evaluated were set into a field which was within 20 feet of a large block of tomatoes which was a continuous source of viruliferous whiteflies throughout the season. No imidacloprid was applied to the plants being evaluated but attempts were made to keep whitefly populations below a threshold which would result in irregular ripening of the fruit (20 immature whitefly / 10 terminal leaflets). Whitefly populations were evaluated approximately every 2 week beginning about 4 week after transplanting. Whitefly populations varied each season, with the highest populations occurring in the Fall 1998 trial. The trials consisted of 15 plants per block, with three replications, in a randomized complete block design. Plants were evaluated every other week for the presence of whiteflies and virus. Plants displaying virus-like symptoms were assayed by nucleic...

example 3

Yields

[0031] Results are shown in Tables 1, 2 and 3. The transformed lines yielded as much or more than the untransformed parents and the commercial hybrid ‘Agriset’ in all three trials. The best transformed lines, 02, 04, 11 and 12 yielded approximately 50%-100% more marketable fruit than the untransformed lines. Yields of these transformed lines in the presence of ToMoV and whiteflies were comparable to yields of the untransformed lines in the absence of virus and whiteflies. In addition, transformed plants yielded well in both fall and spring production seasons.

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Abstract

The subject invention pertains to materials and methods for producing plants that are resistant to infection by geminiviruses and other related viruses. Methods of the invention comprise transforming a plant with a polynucleotide wherein when the polynucleotide is expressed in the plant, the transformed plant exhibits resistance to plant viral infections. Exemplified herein is the use of a polynucleotide encoding a Rep protein derived from tomato mottle geminivirus. The methods of the invention can be used to provide resistance to viral infection in plants such as tomato and tobacco. The present invention also concerns transformed and transgenic plants in plant tissue that express a polynucleotide encoding a plant virus Rep protein, or a fragment or variant thereof.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of U.S. application Ser. No. 09 / 491,063, filed Jan. 25, 2000, which claims the benefit of U.S. Provisional Application No. 60 / 117,151, filed Jan. 25, 1999.[0002] The subject invention was made with government support under a research project supported by USDA Grant No. 92341357456 and USDA Grant No. 98341356784. The government has certain rights in this invention.BACKGROUND OF THE INVENTION [0003] Whitefly-transmitted geminiviruses have become a major limiting factor in tomato production in Florida, the Caribbean and much of Latin America. This group of viruses is currently expanding in the Western Hemisphere, and the number of characterized geminiviruses which infect tomato in this region has increased from three to more than 17 over the last 10 years (Polston and Anderson, 1997). This expansion is continuing and reports of new epidemics are appearing almost monthly. Whitefly-transmitted viruses appea...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/01C12N15/34C12N15/82
CPCC07K14/005C12N2750/12022C12N15/8283
Inventor POLSTON, JANEHIEBERT, ERNESTABOUZID, AHMEDHUNTER, WAYNE
Owner UNIV OF FLORIDA RES FOUNDATION INC
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